This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Zhang, J Articles by Aronson, A I Search for Related Content PubMed PubMed Citation Articles by Zhang, J Articles by Aronson, A I

research-article

Cloning and characterization of a cluster of genes encoding polypeptides present in the insoluble fraction of the spore coat of Bacillus subtilis.

Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907.

ABSTRACT

The Bacillus subtilis spore coat is composed of at least 15 polypeptides plus an insoluble protein fraction arranged in three morphological layers. The insoluble fraction accounts for about 30% of the coat protein and is resistant to solubilization by a variety of reagents, implying extensive cross-linking. A dodecapeptide was purified from this fraction by formic acid hydrolysis and reverse-phase high-performance liquid chromatography. This peptide was sequenced, and a gene designated cotX was cloned by reverse genetics. The cotX gene encoding the dodecapeptide at its amino end was clustered with four other genes designated cotV, cotW, cotY, and cotZ. These genes were mapped to 107 degrees between thiB and metA on the B. subtilis chromosome. The deduced amino acid sequences of the cotY and cotZ genes are very similar. Both proteins are cysteine rich, and CotY antigen was present in spore coat extracts as disulfide cross-linked multimers. There was little CotX antigen in the spore coat soluble fraction, and deletion of this gene resulted in a 30% reduction in the spore coat insoluble fraction. Spores produced by strains with deletions of the cotX, cotYZ, or cotXYZ genes were heat and lysozyme resistant but readily clumped and responded more rapidly to germinants than did spores from the wild type. In electron micrographs, there was a less densely staining outer coat in spores produced by the cotX null mutant, and those produced by a strain with a deletion of the cotXYZ genes had an incomplete outer coat. These proteins, as part of the coat insoluble fraction, appear to be localized to the outer coat and influence spore hydrophobicity as well as the accessibility of germinants.

This article has been cited by other articles:

• Johnson, M. J., Todd, S. J., Ball, D. A., Shepherd, A. M., Sylvestre, P., Moir, A. (2006). ExsY and CotY Are Required for the Correct Assembly of the Exosporium and Spore Coat of Bacillus cereus. J. Bacteriol. 188: 7905-7913 [Abstract] [Full Text]  
• Monroe, A., Setlow, P. (2006). Localization of the Transglutaminase Cross-Linking Sites in the Bacillus subtilis Spore Coat Protein GerQ. J. Bacteriol. 188: 7609-7616 [Abstract] [Full Text]  
• Klobutcher, L. A., Ragkousi, K., Setlow, P. (2006). The Bacillus subtilis spore coat provides "eat resistance" during phagocytic predation by the protozoan Tetrahymena thermophila. Proc. Natl. Acad. Sci. USA 103: 165-170 [Abstract] [Full Text]  
• McPherson, D. C., Kim, H., Hahn, M., Wang, R., Grabowski, P., Eichenberger, P., Driks, A. (2005). Characterization of the Bacillus subtilis Spore Morphogenetic Coat Protein CotO. J. Bacteriol. 187: 8278-8290 [Abstract] [Full Text]  
• Zilhao, R., Isticato, R., Martins, L. O., Steil, L., Volker, U., Ricca, E., Moran, C. P. Jr, Henriques, A. O. (2005). Assembly and Function of a Spore Coat-Associated Transglutaminase of Bacillus subtilis. J. Bacteriol. 187: 7753-7764 [Abstract] [Full Text]  
• Steil, L., Serrano, M., Henriques, A. O., Volker, U. (2005). Genome-wide analysis of temporally regulated and compartment-specific gene expression in sporulating cells of Bacillus subtilis. Microbiology 151: 399-420 [Abstract] [Full Text]  
• Ragkousi, K., Setlow, P. (2004). Transglutaminase-Mediated Cross-Linking of GerQ in the Coats of Bacillus subtilis Spores. J. Bacteriol. 186: 5567-5575 [Abstract] [Full Text]  
• Kim, H.-S., Sherman, D., Johnson, F., Aronson, A. I. (2004). Characterization of a Major Bacillus anthracis Spore Coat Protein and Its Role in Spore Inactivation. J. Bacteriol. 186: 2413-2417 [Abstract] [Full Text]  
• Zilhao, R., Serrano, M., Isticato, R., Ricca, E., Moran, C. P. Jr., Henriques, A. O. (2004). Interactions among CotB, CotG, and CotH during Assembly of the Bacillus subtilis Spore Coat. J. Bacteriol. 186: 1110-1119 [Abstract] [Full Text]  
• Liu, H., Bergman, N. H., Thomason, B., Shallom, S., Hazen, A., Crossno, J., Rasko, D. A., Ravel, J., Read, T. D., Peterson, S. N., Yates, J. III, Hanna, P. C. (2004). Formation and Composition of the Bacillus anthracis Endospore. J. Bacteriol. 186: 164-178 [Abstract] [Full Text]  
• Ragkousi, K., Eichenberger, P., van Ooij, C., Setlow, P. (2003). Identification of a New Gene Essential for Germination of Bacillus subtilis Spores with Ca2+-Dipicolinate. J. Bacteriol. 185: 2315-2329 [Abstract] [Full Text]  
• Lai, E.-M., Phadke, N. D., Kachman, M. T., Giorno, R., Vazquez, S., Vazquez, J. A., Maddock, J. R., Driks, A. (2003). Proteomic Analysis of the Spore Coats of Bacillus subtilis and Bacillus anthracis. J. Bacteriol. 185: 1443-1454 [Abstract] [Full Text]  
• Kuwana, R., Kasahara, Y., Fujibayashi, M., Takamatsu, H., Ogasawara, N., Watabe, K. (2002). Proteomics characterization of novel spore proteins of Bacillus subtilis. Microbiology 148: 3971-3982 [Abstract] [Full Text]  
• Rudner, D. Z., Pan, Q., Losick, R. M. (2002). Evidence that subcellular localization of a bacterial membrane protein is achieved by diffusion and capture. Proc. Natl. Acad. Sci. USA 99: 8701-8706 [Abstract] [Full Text]  
• Kakeshita, H., Oguro, A., Amikura, R., Nakamura, K., Yamane, K. (2000). Expression of the ftsY gene, encoding a homologue of the {alpha} subunit of mammalian signal recognition particle receptor, is controlled by different promoters in vegetative and sporulating cells of Bacillus subtilis. Microbiology 146: 2595-2603 [Abstract] [Full Text]  
• Ichikawa, H., Kroos, L. (2000). Combined Action of Two Transcription Factors Regulates Genes Encoding Spore Coat Proteins of Bacillus subtilis. J. Biol. Chem. 275: 13849-13855 [Abstract] [Full Text]  
• Wade, K. H., Schyns, G., Opdyke, J. A., Moran, C. P. Jr. (1999). A Region of sigma K Involved in Promoter Activation by GerE in Bacillus subtilis. J. Bacteriol. 181: 4365-4373 [Abstract] [Full Text]  
• Serrano, M., Zilhão, R., Ricca, E., Ozin, A. J., Moran, C. P. Jr., Henriques, A. O. (1999). A Bacillus subtilis Secreted Protein with a Role in Endospore Coat Assembly and Function. J. Bacteriol. 181: 3632-3643 [Abstract] [Full Text]  
• Driks, A. (1999). Bacillus subtilis Spore Coat. Microbiol. Mol. Biol. Rev. 63: 1-20 [Abstract] [Full Text]  
• Takamatsu, H., Chikahiro, Y., Kodama, T., Koide, H., Kozuka, S., Tochikubo, K., Watabe, K. (1998). A Spore Coat Protein, CotS, of Bacillus subtilis Is Synthesized under the Regulation of sigma K and GerE during Development and Is Located in the Inner Coat Layer of Spores. J. Bacteriol. 180: 2968-2974 [Abstract] [Full Text]  
• Henriques, A. O., Melsen, L. R., Moran, C. P. Jr. (1998). Involvement of Superoxide Dismutase in Spore Coat Assembly in Bacillus subtilis. J. Bacteriol. 180: 2285-2291 [Abstract] [Full Text]